Ureylenebis(anionic substituted phenylene carbonyl)imino naphthalene sulfonic acids and naphthalene carboxylic acids and their salts

ABSTRACT

Ureylenebis(anionic substituted phenylene carbonyl)-imino naphthalene sulfonic acids and naphthalene carboxylic acids and their salts, which are useful as inhibitors of the complement system of warm-blooded animals. Naphthyl sulfonbenzamido salts, which are new intermediates for the preparation of the active ureylenes, and the process for their preparation.

DESCRIPTION OF THE INVENTION

This invention is concerned with naphthalene sulfonic acid andnaphthalene carboxylic acid ureylenes and their pharmaceuticallyacceptable salts, having complement inhibiting activity, which are newcompounds of the general formula (I): ##STR1## wherein R is selectedfrom the group consisting of SO₃ X and COOY, wherein X is selected fromthe group consisting of alkali metal and Y is selected from the groupconsisting of hydrogen, alkali metal and C₁ -C₆ alkyl; R₁, R₄, R₅, andR₆ are selected from the group consisting of hydrogen and SO₃ X, whereinX is as previously defined; R₂ is selected from the group consisting ofhydrogen and acetamido; with the proviso that there is no R₁ or R₂substituent when the bridgehead carbonylimino is attached at the carbon2-position of the respective ring; with the further proviso that eachnaphthalene moiety must contain two or three SO₃ X substituents, whereinX is as previously defined; R₃ is selected from the group consisting ofhydrogen, SO₃ X and COOY, wherein X and Y are as previously defined; R₇is selected from the group consisting of hydrogen, hydroxy and SO₃ X,wherein X is as previously defined; and the pharmaceutically acceptablesalts thereof.

A preferred embodiment consists of those compounds wherein R, R₁, R₂,R₃, R₄, R₅, and R₆ are as previously defined; and R₇ is selected fromthe group consisting of SO₃ X, wherein X is as previously defined.

A second preferred embodiment consists of those compounds wherein R, R₁,R₂, R₃, R₄, R₅ and R₆ are as previously defined; and R₇ is selected fromthe group consisting of hydroxy and hydrogen.

A third preferred embodiment consists of those compounds wherein R₁, R₂,R₃, R₄, R₅, R₆ and R₇ are as previously defined; and R is selected fromthe group consisting of SO₃ X, wherein X is as previously defined.

A fourth preferred embodiment consists of those compounds wherein R₁,R₂, R₃, R₄, R₅, R₆ and R₇ are as previously defined; and R is selectedfrom the group consisting of COOY, wherein Y is as previously defined.

A most preferred embodiment of the third preferred embodiment consistsof those compounds wherein R, R₁, R₃, R₄, R₅, R₆ and R₇ are aspreviously defined; and R₂ is hydrogen.

A further preferred embodiment of the above most preferred embodimentconsists of those compounds wherein R, R₁, R₂, R₃, R₄, R₅ and R₆ are aspreviously defined; and R₇ is selected from the group consisting ofhydrogen and hydroxy.

A second further preferred embodiment of the above most preferredembodiment consists of those compounds wherein R, R₁, R₂, R₃, R₄, R₅ andR₆ are as previously defined; and R₇ is selected from the groupconsisting of SO₃ X, wherein X is as previously defined.

A most preferred embodiment of fourth preferred embodiment consists ofthose compounds wherein R, R₁, R₃, R₄, R₅, R₆ and R₇ are as previouslydefined; and R₂ is hydroxy.

A further preferred embodiment of the above most preferred embodimentconsists of those compounds wherein R, R₁, R₂, R₃, R₄, R₅ and R₆ are aspreviously defined; and R₇ is selected from the group consisting ofhydrogen and hydroxy.

A second further preferred embodiment of the above most preferredembodiment consists of those compounds wherein R, R₁, R₂, R₃, R₄, R₅ areas previously defined; and R₇ is selected from the group consisting ofSO₃ X, wherein X is as previously defined.

This invention is also concerned with compounds of the formula (II):##STR2## wherein R is selected from the group consisting of SO₃ X andCOOY, wherein X is selected from the group consisting of hydrogen andalkali metal and Y is selected from the group consisting of hydrogen,alkali metal and C₁ -C₆ alkyl; R₁, R₄, R₅ and R₆ are selected from thegroup consisting of hydrogen and SO₃ X, wherein X is as previouslydefined; R₂ is selected from the group consisting of hydrogen andacetamido; with the proviso that there is no R₁ or R₂ substituent whenthe bridgehead carbonylimino is attached at the carbon 2-position of therespective ring; R₃ is selected from the group consisting of hydrogen,SO₃ X and COOY, wherein X and Y are as previously defined; and R₇ isselected from the group consisting of hydrogen, hydroxy and SO₃ X,wherein X is as previously defined.

A preferred embodiment consists of those compounds wherein R, R₁, R₂,R₃, R₄, R₅ and R₆ are as previously defined; and R₇ is selected from thegroup consisting of SO₃ X, wherein X is as previously defined.

A second preferred embodiment consists of those compounds wherein R, R₁,R₂, R₃, R₄, R₅ and R₆ are as previously defined; and R₇ is selected fromthe group consisting of hydroxy and hydrogen.

A third preferred embodiment consists of those compounds, wherein R₁,R₂, R₃, R₄, R₅, R₆ and R₇ are as previously defined; and R is selectedfrom the group consisting of SO₃ X, wherein X is as previously defined.

A fourth preferred embodiment consists of those compounds wherein R₁,R₂, R₃, R₄, R₅, R₆ and R₇ are as previously defined; and R is selectedfrom the group consisting of COOY, wherein Y is as previously defined.

A most preferred embodiment of the third preferred embodiment consistsof those compounds wherein R, R₁, R₃, R₄, R₅, R₆ and R₇ are aspreviously defined; and R₂ is hydrogen.

A further preferred embodiment of the above most preferred embodimentconsists of those compounds wherein R, R₁, R₂, R₃, R₄, R₅ and R₆ are aspreviously defined; and R₇ is selected from the group consisting ofhydrogen and hydroxy.

A second further preferred embodiment of the above most preferredembodiment consists of those compounds wherein R, R₁, R₂, R₃, R₄, R₅ andR₆ are as previously defined; and R₇ is selected from the groupconsisting of SO₃ X, wherein X is as previously defined.

A most preferred embodiment of fourth preferred embodiment consists ofthose compounds wherein R, R₁, R₃, R₄, R₅, R₆ and R₇ are as previouslydefined; and R₂ is hydrogen.

A further preferred embodiment of the above most preferred embodimentconsists of those compounds wherein R, R₁, R₂, R₃, R₄, R₅ and R₆ are aspreviously defined; and R₇ is selected from the group consisting ofhydrogen and hydroxy.

A second further preferred embodiment of the above most preferredembodiment consists of those compounds wherein R, R₁, R₂, R₃, R₄, R₅ areas previously defined; and R₇ is selected from the group consisting ofSO₃ X, wherein X is as previously defined.

These compounds are useful as intermediates for the preparation of thecomplement inhibiting compounds described above. The compounds of thepresent invention may be prepared by the following method outlined inFlow Chart A. ##STR3## wherein R, R₁, R₂, R₃, R₄, R₅, R₆ and R₇ are aspreviously defined.

The novel intermediate amine compounds (III) of the invention areprepared by reacting the appropriate naphthylamine with a 15-20% excessof 3- or 4-nitro-2-sulfobenzoic anhydride in aqueous media with sodiumacetate as a buffer at 0°-5° C. for approximately 10 minutes.Acidification of the mixture to approximately pH 1.0 followed bydilution with ethyl alcohol to approximately 80% aqueous ethyl alcoholresults in precipitation of the nitroamide (II). Hydrogenation of thenitroamide using 10% palladium-carbon catalyst provides the intermediateamine (III).

The novel ureylene compounds (IV) of the invention, which are activecomplement inhibitors, are then provided by treatment of the amine (III)with phosgene in aqueous medium containing sodium carbonate.

A method of preparing the nitroamide,8-(p-nitro-sulfobenzamido)-1,3,6-naphthalenetrisulfonic acid trisodiumsalt (IIa), as described in Flow Chart B, involves the reaction of8-amino-1,3,6-naphthalenetrisulfonic acid trisodium salt (Ia) andp-nitrosulfobenzoic anhydride in base. The subsequent acidification andextraction of the aqueous solution with diethyl ether, followed byconcentration and crystallization of the nitroamide (IIa) from theaqueous medium. The nitroamide is then hydrogenated and the amine (IIIa)is phosgenated to yield the product8,8'-{ureylenebis[(2-sulfo-4,1-phenylenecarbonyl)imino]}di-1,3,6-naphthalenetrisulfonicacid hexasodium salt (IVa). ##STR4##

Alternatively, the reaction of 8-amino-1,3,6-naphthalenetrisulfonic acidtrisodium salt and 8-amino-1,3,5-naphthalenetrisulfonic acid trisodiumsalt, respectively, with 5-nitroisophthaloyl chloride and3-carbomethoxy-5-nitrobenzoyl chloride, respectively, in aqueous mediumwith sodium acetate as buffer, results in the recovery of thenitroamides 5-nitro-N-3,6,8-trisulfo-1-naphthylisophthalamic acidtrisodium salt and 5-nitro-N-4,6,8-trisulfo-1-naphthylisophthalamic acidmethyl ester trisodium salt, respectively, by crystallization from ethylalcohol. Hydrogenation of the respective nitroamide provides thecorresponding amine (III) which in turn is phosgenated to yield therespective novel ureide compound (IV).

The term "complement" refers to a complex group of proteins in bodyfluids that, working together with antibodies or other factors, play animportant role as mediators of immune, allergic, immunochemical and/orimmunopathological reactions. The reactions in which complementparticipates take place in blood serum or in other body fluids, andhence are considered to be humoral reactions.

With regard to human blood, there are at present more than 11 proteinsin the complement system. These complement proteins are designated bythe letter C and by number: C1, C2, C3 and so on up to C9. Thecomplement protein C1 is actually an assembly of subunits designatedC1q, C1r and C1s. The numbers assigned to the complement proteinsreflect the sequence in which they become active, with the exception ofcomplement protein C4, which reacts after C1 and before C2. Thenumerical assignments for the proteins in the complement system weremade before the reaction sequence was fully understood. A more detaileddiscussion of the complement system and its role in body processes canbe found in, for example, Bull. World Health Org., 39, 935-938 (1968);Ann. Rev. Medicine, 19, 1-24 (1968); The John Hopkins Med. J., 128,57-74 (1971); Harvey Lectures, 66, 75-104 (1972); The New EnglandJournal of Medicine, 287, 452-454; 489-495; 545-549; 592-596; 642-646(1972); Scientific American, 229, (No. 5), 54-66 (1973); FederationProceedings, 32, 134-137 (1973); Medical World News, October 11, 1974,pp. 53-58; 64-66; J. Allergy Clin. Immunol., 53, 298-302 (1974); ColdSpring Harbor Conf. Cell Proliferation 2/Proteases Biol. Control/229-241(1975); Annals of Internal Medicine, 84, 580-593 (1976); "Complement:Mechanisms and Functions", Prentice-Hall, Englewood Cliffs, N. J.(1976).

The complement system can be considered to consist of three sub-systems:(1) a recognition unit (C1q) which enables it to combine with antibodymolecules that have detected a foreign invader; (2) an activation unit(C1r, C1s, C2, C4, C3) which prepares a site on the neighboringmembrane; and (3) and attack unit (C5, C6, C7, C8 and C9) which createsa "hole" in the membrane. The membrane attack unit is non-specific; itdestroys invaders only because it is generated in their neighborhood. Inorder to minimize damage to the host's own cells, its activity must belimited in time. This limitation is accomplished partly by thespontaneous decay of activated complement and partly by interference byinhibitors and destructive enzymes. The control of complement, however,is not perfect, and there are times when damage is done to the host'scells. Immunity is therefore a double-edged sword.

Activation of the complement system also accelerates blood clotting.This action comes about by way of the complement-mediated release of aclotting factor from platelets. The biologically active complementfragments and complexes can become involved in reactions that damage thehost's cells, and these pathogenic reactions can result in thedevelopment of immune-complex diseases. For example, in some forms ofnephritis, complement damages the basal membrane of the kidney,resulting in the escape of protein from the blood into the urine. Thedisease disseminated lupus erythematosus belongs in this category; itssymptoms include nephritis, visceral lesions and skin eruptions. Thetreatment of diphtheria or tetanus with the injection of large amountsof antitoxin sometimes results in serum sickness, an immune-complexdisease. Rheumatoid arthritis also involves immune complexes. Likedisseminated lupus erythematosus, it is an autoimmune disease in whichthe disease symptoms are caused by pathological effects of the immunesystem in the host's tissues. In summary, the complement system has beenshown to be involved with inflammation, coagulation, fibrinolysis,antibody-antigen reactions and other metabolic processes.

In the presence of antibody-antigen complexes the complement proteinsare involved in a series of reactions which may lead to irreversiblemembrane damage if they occur in the vicinity of biological membranes.Thus, while complement constitutes a part of the body's defensemechanism against infection it also results in inflammation and tissuedamage in the immunopathological process. The nature of certain of thecomplement proteins, suggestions regarding the mode of complementbinding to biological membranes and the manner in which complementeffects membrane damage are discussed in Annual Review in Biochemistry,38, 389 (1969).

A variety of substances have been disclosed as inhibiting the complementsystem, i.e., as complement inhibitors. For example, the compounds3,3'-ureylenebis-[6-(2-amino-8-hydroxy-6-sulfo-1-naphthylazo)]benzenesulfonicacid, tetrasodium salt (chlorazol fast pink), heparin and a sulphateddextran have been reported to have an anticomplementary effect, BritishJournal of Experimental Pathology, 33, 327-339 (1952). The compound8-(3-benzamido-4-methylbenzamido)naphthalene-1,3,5-trisulfonic acid(Suramin) is described as a competitive inhibitor of the complementsystem, Clin. Exp. Immunol., 10, 127-138 (1972). German Patent No.2,254,893 or South African Patent No. 727,923 discloses certain1-(diphenylmethyl)-4-(3-phenylallyl)piperazines useful as complementinhibitors. Other chemical compounds having complement inhibitingactivity are disclosed in, for example, Journal of Medicinal Chemistry,12, 415-419; 902-905; 1049-1052; 1053-1056 (1969); Canadian Journal ofBiochemistry, 47, 547-552 (1969); The Journal of Immunology, 93, 629-640(1964); The Journal of Immunology, 104, 279-288 (1970); The Journal ofImmunology, 106, 241-245 (1971); and The Journal of Immunology, 111,1061-1066 (1973).

It has been reported that the known complement inhibitorsepsilon-aminocaproic acid, Suramin and tranexamic acid all have beenused with success in the treatment of hereditary angioneurotic edema, adisease state resulting from an inherited deficiency or lack of functionof the serum inhibitor of the activated first component of complement(C1 inhibitor), The New England Journal of Medicine, 286, 808-812(1972).

The compounds of the present invention may be administered internally,e.g., orally, intra-articularly or parenterally, e.g., intra-articular,to a warm-blooded animal to inhibit complement in the body fluid of theanimal, such inhibition being useful in the amelioration or preventionof those reactions dependent upon the function of complement, such asinflammatory process and cell membrane damage induced byantigen-antibody complexes. A range of doses may be employed dependingon the mode of administration, the condition being treated and theparticular compound being used. For example, for intravenous orsubcutaneous use from about 5 to about 50 mg/kg/day, or every six hoursfor more rapidly excreted salts, may be used. For intra-articular usefor large joints such as the knee, from about 2 to about 20 mg/joint perweek may be used, with proportionally smaller doses for smaller joints.The dosage range is to be adjusted to provide optimum therapeuticresponse in the warm-blooded animal being treated. In general, theamount of compound administered can vary over a wide range to providefrom about 5 mg/kg to about 100 mg/kg of body weight of animal per day.The usual daily dosage for a 70 kg subject may vary from about 350 mg toabout 3.5 g. Unit doses of the acid or salt can contain from about 0.5mg to about 500 mg.

While in general the sodium salts of the acids of the invention aresuitable for parenteral use, other salts may also be prepared, such asthose of primary amines, e.g., ethylamine; secondary amines, e.g.,diethylamine or diethanol amine; tertiary amines, e.g., pyridine ortriethylamine or 2-dimethylaminomethyl-dibenzofuran; aliphatic diamines,e.g., decamethylenediamine; and aromatic diamines, can be prepared. Someof these are soluble in water, others are soluble in saline solution,and still others are insoluble and can be used for purposes of preparingsuspensions for injection. Furthermore as well as the sodium salt, thoseof the alkali metals, such as potassium and lithium; of ammonia; and ofthe alkaline earth metals, such as calcium or magnesium, may beemployed. It will be apparent, therefore, that these salts embrace, ingeneral derivatives of salt-forming cations.

In therapeutic use, the compounds of this invention may be administeredin the form of conventional pharmaceutical compositions. Suchcompositions may be formulated so as to be suitable for oral orparenteral administration. The active ingredient may be combined inadmixture with a pharmaceutically acceptable carrier, which carrier maytake a wide variety of forms depending on the form of preparationdesired for administration, i.e., oral or parenteral. The compounds canbe used in compositions such as tablets. Here, the principal activeingredient is mixed with conventional tabletting ingredients such ascorn starch, lactose, sucrose, sorbitol, talc, stearic acid, magnesiumstearate, dicalcium phosphate, gums, or similar materials as non-toxicpharmaceutically acceptable diluents or carriers. The tablets or pillsof the novel compositions can be laminated or otherwise compounded toprovide a dosage form affording the advantage of prolonged or delayedaction or predetermined successive action of the enclosed medication.For example, the tablet or pill can comprise an inner dosage and anouter dosage component, the latter being in the form of an envelope overthe former. The two components can be separated by an enteric layerwhich serves to resist disintegration in the stomach and permits theinner component to pass intact into the duodenum or to be delayed inrelease. A variety of materials can be used for such enteric layers orcoatings, such materials including a number of polymeric acids ormixtures of polymeric acids with such materials as shellac, shellac andcetyl alcohol, cellulose acetate and the like. A particularlyadvantageous enteric coating comprises a styrene maleic acid copolymertogether with known materials contributing to the enteric properties ofthe coating. The tablet or pill may be colored through the use of anappropriate nontoxic dye, so as to provide a pleasing appearance.

The liquid forms in which the novel compositions of the presentinvention may be incorporated for administration include suitableflavored emulsions with edible oils, such as, cottonseed oil, sesameoil, coconut oil, peanut oil, and the like, as well as elixirs andsimilar pharmaceutical vehicles. Sterile suspensions or solutions can beprepared for parenteral use. Isotonic preparations containing suitablepreservatives are also desirable for injection use.

The term dosage form, as described herein, refers to physically discreteunits suitable as unitary dosage for warm-blooded animal subjects, eachunit containing a predetermined quantity of active component calculatedto produce the desired therapeutic effect in association with therequired pharmaceutical diluent, carrier or vehicle. The specificationfor the novel dosage forms of this invention are indicated bycharacteristics of the active component and the particular therapeuticeffect to be achieved or the limitations inherent in the art ofcompounding such an active component for therapeutic use in warm-bloodedanimals as disclosed in this specification. Examples of suitable oraldosage forms in accord with this invention are tablets, capsules, pills,powder packets, granules, wafers, cachets, teaspoonfuls, dropperfuls,ampules, vials, segregated multiples of any of the foregoing and otherforms as herein described.

The complement inhibiting activity of the compounds of this inventionhas been demonstrated by one or more of the following identified tests:(i) Test, Code 026 (C1 inhibitor) - This test measures the ability ofactivated human C1 to destroy fluid phase human C2 in the presence of C4and appropriate dilutions of the test compound. An active inhibitorprotects C2 from C1 and C4; (ii) Test, Code 035 (C3-C9 inhibitor) - Thistest determines the ability of the late components of human complement(C3-C9) to lyse EAC 142 in the presence of appropriate dilutions of thetest compound. An active inhibitor protects EAC 142 from lysis by humanC3-C9; (iii) Test, Code 036 (C-Shunt inhibitor) - In this test humanerythrocytes rendered fragile are lysed in autologous serum via theshunt pathway activated by cobra venom factor in the presence ofappropriate dilutions of the test compound. Inhibition of the shuntpathway results in failure of lysis; (iv) Forssman Vasculitis Test -Here, the well known complement dependent lesion, Forssman vasculitis,is produced in guinea pigs by intradermal injection of rabbitanti-Forssman antiserum. The lesion is measured in terms of diameter,edema and hemorrhage and the extent to which a combined index of theseis inhibited by prior intraperitoneal injection of the test compound at200 mg/kg is then reported, unless otherwise stated; (v) Forssman ShockTest - Lethal shock is produced in guinea pigs by an i.v. injection ofanti-Forssman antiserum and the harmonic mean death time of treatedguinea pigs is compared with that of simultaneous controls; (vi)Complement Level Reduction Test - In this test, the above dosed guineapigs, or others, are bled for serum and the complement level isdetermined in undiluted serum by the capillary tube method of U.S.Patent No. 3,876,376 and compared to undosed control guinea pigs; and(vii) Cap 50 Test - Here, appropriate amounts of the test compound areadded to a pool of guinea pig serum in vitro, after which the undilutedserum capillary tube assay referred to above is run. The concentrationof compound inhibiting 50% is reported.

With reference to Table I, guinea pigs weighing about 300 g were dosedintravenously (i.v.) or intraperitoneally (i.p.) with 200 mg/kg of thetest compound dissolved in saline and adjusted to pH 7-8. One hour afterdosing, the guinea pigs were decapitated, blood was collected and theserum separated. The serum was tested for whole complement using thecapillary tube assay. Percent inhibition was calculated by comparisonwith simultaneous controls. The results appear in Table I together withresults of tests, Code 026, 035, 036, Cap 50, percent inhibition andForssman shock. Table I shows that the compounds of the inventionpossess highly significant in vitro and in vivo, complement inhibitingactivity in warm-blooded animals.

                                      TABLE I                                     __________________________________________________________________________    Biological Activities                                                                                                In Vivo Activity (Guinea Pig)                                                 % Inhibition                                               Cl  C-Late                                                                            Shunt Inhi-                                                                              Intraperitoneal                                                                        Intravenous                                       026*                                                                              035*                                                                              bition 036*                                                                              Time (mins.)                                                                           Time (mins.)                  Compound            Wells                                                                             Wells                                                                             Wells Cap 50*                                                                            30 60 120                                                                              2  30 120                     __________________________________________________________________________    8,8'-[Ureylenebis(4,1-phenylenecarbo-                                                             +4**                                                                              +1  +3                                                nylimino)]di-1,3,6-naphthalenetri-                                                                +4  N   N     290  -18                                                                              -21                                                                              -33                                                                              -71                                                                              -28                                                                              -16                     sulfonic acid hexasodium salt                                                 8,8'-{Ureylenebis{[(2-sulfo-p-pheny-                                                              +7  +1  +3                                                lene)carbonyl]imino}}-1,3,6-naphtha-                                                              +8  N   N                                                 lenetrisulfonic acid octasodium salt                                                              +8  +1  +4    130  +4 +17                                                                              -8 -85                                                                              -53                                                                              -24                     5,5'-[Ureylenebis(2-sulfo-4,1-pheny-                                                              +8  N   +1    305  -19                                                                              +8 +4                               lenecarbonylimino)]bis[6-acetamido-                                           1,3-naphthalenedisulfonic acid]                                               hexasodium salt                                                               4,4'-{Ureylenebis[(2-sulfo-4,1-                                                                   +9  +1  + 3   194  -35                                                                              -41                                                                              -74                                                                              -98                                                                              -81                                                                              -63                     phenylenecarbonyl)imino]}bis-[5-                                                                  +6  +1  +2                                                hydroxy-2,7-naphthalenedisulfonic                                                                 +6  +2  +4     80                                         acid]hexasodium salt                                                          8,8'-{Ureylenebis[(2-sulfo-4,1-pheny-                                                             +8  +1  +1                                                lenecarbonyl)imino]}di-1,3,5-naph-                                                                +7  +1  +3    104  -32                                                                              -31                                                                              -51                                                                              -89                                                                              -45                                                                              -29                     thalenetrisulfonic acid octasodium                                                                +8  +1  +3                                                salt                                                                          3,3'-{Ureylenebis[(2-sulfo-4,1-phen-                                                              N   +3  N     >500                                        ylene)carbonylimino]}di-2-naphthoic                                           acid tetrasodium salt                                                         4,4'-{Ureylenebis[(2-sulfo-4,1-phen-                                                              +4  N   N                                                 ylenecarbonyl)imino]}di-2,7-naphtha-                                                              +5  +1  N     >500                                        lenedisulfonic acid hexasodium salt                                           5,5'-{Ureylenebis[(2-sulfo-4,1-phen-                                                              +2  N   +1                                                ylenecarbonyl)imino]}bis[4-hydroxy-                                                               +2  +1  N     102  -26                                                                              0  -58                              2-naphthalenesulfonic acid] tetra-                                            sodium salt                                                                   4,4'-Ureylenebis[(2-sulfo-4,1-phen-                                                               +7  N   +5    <100 +1 -3 -24                              ylenecarbonyl)imino]bis[5-hydroxy-                                            1,7-naphthalenedisulfonic acid]                                               hexasodium salt                                                               8,8'-[Ureylenebis(2-sulfo-4,1-phen-                                                               +5  +1  +1    297  -10                                                                              -18                                                                              -31                              ylenecarbonylimino)]di-1,6-naphtha-                                           lenedisulfonic acid hexasodium salt                                           4,4'-{Ureylenebis[(2-sulfo-4,1-phen-                                                              +5  N   N     277  - 3                                                                              -4 +6                               ylenecarbonyl)imino]}di-1,6-naphtha-                                          lenedisulfonic acid hexasodium salt                                           4,4'-{Ureylenebis[(2-sulfo-4,1-phen-                                                              +5  N   N     170  -3 -4 -11                              ylenecarbonyl)imino]}di-2,6-naphtha-                                          lenedisulfonic acid hexasodium salt                                           4,4'-{Ureylenebis[(2-sulfo-4,1-phen-                                                              +7  N   +1         -10                                                                              +6 -6                               ylenecarbonyl)imino]}di-1,5-naphtha-                                          lenedisulfonic acid trisodium salt                                            5,5'-Ureylenebis[N-(3,6,8-trisulfo-                                                               +10 N   +2    146  -37                                                                              -42                                                                              -48                                                                              -92                                                                              -44                                                                              -39                     1-naphthyl)]isophthalamic acid                                                octasodium salt                                                               5,5'-Ureylenebis[N-(4,6,8-trisulfo-                                                               +4  +1  +1    249  -14                                                                              -17                                                                              -25                              1-naphthyl)isophthalamic acid]di-                                             methyl ester hexasodium salt                                                  5,5'-Ureylenebis[N-(4,6,8-trisulfo-                                                               +5  +1  +2    <100 -40                                                                              -49                                                                              -71                              1-naphthyl)isophthalamic acid]oc-                                             tasodium salt                                                                 3,3'-Ureylenebis{5-[(8-hydroxy-4,6-                                                               +4  +2  +2    129                                         disulfo-1-naphthyl)aminocarbonyl]-                                                                +4  +4                                                    benzoic acid}tetrasodium salt                                                 4,4'-}Ureylenebis[(6-methyl-3,1-                                                                  +4  +1  +2                                                phenylenecarbonyl)imino]}bis[5-                                                                   +8  +3                                                    hydroxy-1,7-naphthalenedisulfonic                                             acid],tetrasodium salt                                                        8,8'-[Carbonylbis[(imino-2-sulfo-                                                                 +7  +1  +1    <100 -23                                                                              -58                                                                              -71                              3,1-phenylene)carbonylimino]]bis-                                                                 +4  +4                                                    1,3,6-naphtholenetrisulfonic acid                                             octosodium salt                                                               8,8'-Carbonylbis[(imino-6-sulfo-                                              3,1-phenylene)carbonylimino]}bis-                                                                 +7            48         -71                              1,3,6-naphthalenetrisulfonic acid                                             octasodium salt                                                               3,3-(Carbonyldiimino)bis{5-[(8-                                               hydroxy-3,6-disulfo-1-naphthalen-                                                                 +6            56         -69                              yl)aminocarbonyl]}benzoic acid                                                tetrasodium salt                                                              3,3'-Ureylenebis{5-[(8-hydroxy-4,6-                                           disulfo-1-naphthyl)aminocarbonyl]}-                                                               +4            129        -94                              benzoic acid tetrasodium salt                                                 __________________________________________________________________________     N=Negative                                                                    *Code designation for tests employed as referred to herein.                   **Activity in wells, a serial dilution assay. Higher well number indicate     higher activity. The serial dilutions are two-fold.                      

SPECIFIC DISCLOSURE EXAMPLE 18-(p-Aminobenzamido)-1,3,6-naphthalenetrisulfonic acid trisodium salt

To a solution of 27.0 g of 8-amino-1,3,6-naphthalenetrisulfonic acidtrisodium salt in 100 ml of water and 60 ml of N sodium hydroxide isadded 22.5 g of p-nitrobenzoyl chloride and 30 ml of diethyl ether. Thereaction mixture is shaken in a separatory funnel for about 15 minutes,and another 60 ml portion of base is added. After shaking for 15 minutesthe procedure is repeated with two additional 60 ml portions of base.The reaction mixture is acidified with 7.5 ml of concentratedhydrochloric acid and diluted with 250 ml of water, then extracted 9times with 300 ml portions of ether. The aqueous phase is neutralizedwith 5N sodium hydroxide and allowed to stand overnight at roomtemperature, then is filtered through diatomaceous earth. The filtrateis concentrated to a low volume in vacuo at 55°-60° C. The crystallinesolid formed is collected by filtration, washed and slurried withabsolute ethyl alcohol to yield 28.6 g of8-(p-nitrobenzamido)-1,3,6-naphthalenetrisulfonic acid trisodium salt.

A mixture of 25.0 g of the above product, 100 ml of water and 2.5 g of10% palladium catalyst on carbon is hydrogenated at room temperature for2 hours and 40 minutes at an average pressure of 37 pounds, then isfiltered through diatomaceous earth and washed with water. The filtrateis concentrated to a small volume in vacuo at 55°-60° C. resulting inthe formation of off-white crystals. The material is diluted with about300 ml of absolute ethyl alcohol, triturated, filtered and washed withabsolute ethanol and dried at 120° C. overnight to give 21.8 g of8-(p-aminobenzamido)-1,3,6-naphthalenetrisulfonic acid trisodium salt asyellow crystals.

EXAMPLE 28,8'-[Ureylenebis(4,1-phenylenecarbonylimino)]di-1,3,6-naphthalenetrisulfonicacid hexasodium salt

Phosgene is bubbled into a solution of 10.0 g of the product of Example1 and 18.6 g of anhydrous sodium carbonate in 250 ml of water until thesolution is acidic. An additional 18.6 g of the sodium carbonate isadded and phosgene is bubbled through for an additional hour. Thereaction mixture is brought to pH 8 by the addition of 75 ml of 5Nsodium hydroxide, then is neutralized with a few drops of 10%hydrochloric acid. The solution is concentrated in vacuo at 50°-55° C.to about 200 ml and is cooled to room temperature. The precipitateformed is collected by filtration and washed with 200 ml of 90% ethylalcohol followed by 250 ml of absolute ethyl alcohol. The product isoven dried at 120° C. overnight, then is slurried with 200 ml of boilingabsolute methyl alcohol and cooled to room temperature. The product ofthe example is collected, washed with cold methyl alcohol and oven driedfor 48 hours at 120° C. to give a colorless powder.

EXAMPLE 3 8-(4-Amino-2-sulfobenzamido)-1,3,6-naphthalenetrisulfonic acidtetrasodium salt

A solution of 100 g of 5-nitro-o-toluenesulfonic acid in 600 ml of waterplus 80 ml of 5N sodium hydroxide is heated to 90° C. in a 2 literErlenmeyer flask, then 240 g of potassium permanganate is addedportionwise to maintain refluxing over 75 minutes. The mixture isfiltered and the residue is washed with water. The combined filtrate andwashings are concentrated in vacuo and allowed to crystallize to give86.2 g of crude 4-nitro-2-sulfobenzoic acid sodium potassium salt.Recrystallization from water gives 71.3 g of purified product.

The total product above is dissolved in 250 ml of water plus 35 ml ofconcentrated hydrochloric acid by warming on a steam-bath. The solutionis then diluted with 300 ml of ethyl alcohol and allowed to crystallizeat room temperature. The mixture is allowed to stand 48 hours in a chillroom, then is filtered. The precipitate is washed with cold 50% aqueousethyl alcohol, then with ethanol and ether. The material isrecrystallized from 200 ml of water and is dried at 110° C. to give 52.0g of 4-nitro-2-sulfobenzoic acid, 2-sodium salt.

A 50.0 g portion of the preceding compound and 500 g of thionyl chlorideare stirred and refluxed for 19 hours. The mixture is evaporated invacuo, and the residue is warmed with 300 ml of toluene and filtered.The filtrate is concentrated in vacuo, and the product is crystallizedtwice from toluene to give 30.4 g of 4-nitro-2-sulfobenzoic acidanhydride.

To an ice-bath cooled solution (5° C.) of8-amino-1,3,6-naphthalenetrisulfonic acid trisodium salt and 6.7 g ofsodium acetate trihydrate in 100 ml of water is added 8.8 g of4-nitro-2-sulfobenzoic acid anhydride. The mixture is stirred vigorouslyfor 10 minutes and is filtered. The filtrate is cooled in an ice bathand diluted with 500 ml of cold ethyl alcohol. The mixture is filteredand the product is washed with ethyl alcohol and ether and then isdried. The product is dissolved in 50 ml of warm water and is stirredfor 10 minutes after the addition of one ml of acetone. The mixture istreated with activated charcoal, filtered through diatomaceous earth andis washed with 20 ml of water. The filtrate is cooled in an ice bath andacidified with 1.5 ml of concentrated hydrochloric acid. The solution isdiluted with 500 ml of cold ethyl alcohol, and the precipitated materialis filtered, washed with ethanol followed by ether and dried. The abovepurification process is repeated without acidification. The productobtained is dried overnight at 110° C. to give 18.5 g of8-(4-nitro-2-sulfobenzamido)-1,3,6-naphthalenetrisulfonic acidtetrasodium salt.

A 17.5 g portion of the preceding product and 1.5 g of palladiumcatalyst on carbon in 150 ml of water is hydrogenated for one hour atroom temperature then is filtered through diatomaceous earth. Thefiltrate is concentrated and the product is precipitated by the additionof absolute ethanol. The product is collected and dried overnight in anabderhalden apparatus at 110° C. to give 14.5 g of8-(4-amino-2-sulfobenzmido)-1,3,6-naphthalenetrisulfonic acidtetrasodium salt as a powder.

EXAMPLE 48,8'-{Ureylenebis{[(2-sulfo-4,1-phenylene)carbonyl]imino}}-1,3,6-naphthalenetrisulfonicacid octasodium salt

Phosgene gas is passed through a solution of 5.5 g of the product ofExample 3 in 40 ml of water and 3.8 ml of pyridine until the solution isacidic. The solution is back neutralized with 0.2 ml of pyridine, thenis poured into 250 ml of absolute ethyl alcohol. A gum is formed whichsolidifies on standing. The gum is ground up and washed on a filter withethyl alcohol and ether. The material is dissolved in 35 ml of water, ismade basic (pH 8-9) with 5N sodium hydroxide and is filtered throughdiatomaceous earth. The filtrate is poured into 350 ml of absolute ethylalcohol and is reacidified with a few drops of acetic acid. The productformed is collected and dried by conventional means to give the productof the Example as a beige powder.

EXAMPLE 56-Acetamido-5-(4-amino-2-sulfobenzamido)-1,3-naphthalenedisulfonic acidtrisodium salt

To a solution of 27.6 g of p-nitroaniline in a mixture of 170 ml ofacetic acid, 50 ml of concentrated hydrochloric acid, 100 ml of waterand 200 ml of crushed ice, cooled in an ice-bath is added 13.8 g ofsodium nitrite in 25 ml of water with stirring [solution (A)]. Stirringis continued during preparation of a solution of 65 g of (about 92.5%)6-amino-1,3-naphthalenedisulfonic acid in 80 ml of 5N sodium hydroxideand 100 ml of water [solution (B)]. A 57 g amount of sodium acetatetrihydrate is added to solution (A) followed by the addition of solution(B). The mixture is heated on a steam bath (60° C.) until dissolved,then is filtered and allowed to crystallize at room temperature. Themixture is then cooled to 10° C. The product is collected and washedwith 300 ml of 20% aqueous solution of sodium acetate trihydratefollowed by ethyl alcohol and ether. The material is dried in vacuo togive 87.0 g of 6-amino-5-(pnitrophenylazo)-1,3-naphthalenedisulfonicacid disodium salt as a crystalline product.

To 300 ml of acetic anhydride cooled in an ice-bath is added portionwise100 g of concentrated sulfuric acid, while keeping the temperature below20° C. To this solution is added, portionwise and with stirring, 50 g ofthe preceding product keeping the temperature below 20° C. The mixtureis stirred at room temperature for 1 hour, then is cooled to about 7° C.and is filtered. The product is washed on the filter with acetone, thenis stirred with two successive 500 ml portions of acetone and isfiltered again and dried on the filter to give 49.4 g of product. A 49.0g portion of this material is dissolved in 300 ml of water and isneutralized to pH 6.7 with 20 g of sodium bicarbonate. The solution isthen acidified to pH 5.5 with acetic acid and is filtered. The filtrateis evaporated in vacuo to about 250 ml, and is poured into 1800 ml ofvigorously stirring absolute ethyl alcohol. The precipitate formed iscollected by filtration and is washed with ethyl alcohol followed byether. A 47.0 g quantity of material dried in vacuo is additionallydried overnight in the Abderhalden apparatus at 110° C. to give 46.5 gof 6-acetamido-5-(p-nitrophenylazo)-1,3-naphthalenedisulfonic aciddisodium salt as a red-brown powder.

A 23.0 g portion of the above product and 1.5 g of palladium catalyst oncarbon in 150 ml of water is hydrogenated in a Parr shaker at roomtemperature for 1.5 hours. The mixture is filtered through diatomaceousearth and the filtrate is concentrated to 80-100 ml. This solution ispoured into 600 ml of ice-cold ethyl alcohol with vigorous stirring in abaffle flask. The mixture is stirred 3-4 minutes then filtered. Theproduct is washed with ethyl alcohol and ether and is dried overnight at65° C. in an Abderhalden apparatus to give 16.15 g of6-acetamido-5-amino-1,3-naphthalenedisulfonic acid disodium salt as apowder.

To a solution of 12.76 g of the preceding compound and 6.25 g of sodiumacetate trihydrate in 75 ml of water, cooled to 3° C. is added 8.25 g of4-nitro-2-sulfobenzoic acid anhydride. The mixture is stirred 10minutes, treated with activated charcoal and filtered throughdiatomaceous earth. The filtrate is concentrated to 50 ml, is cooled inan ice-bath, acidified with 2.5 ml of concentrated hydrochloric acid andpoured with vigorous stirring into 500 ml of ice cold ethyl alcohol. Theresulting gelatinous precipitate is collected by filtration and washedwith ethyl alcohol. The product is stirred in ether, is filtered and isdried in vacuo to yield a powder. The product is further purified byextraction with methanol, filtering and evaporation. The material isdried overnight in an Abderhalden apparatus at 110° C. to yield 6.06 gof 6-acetamido-5-(4-nitro-2-sulfobenzamido)-1,3-naphthalenedisulfonicacid trisodium salt as a powder.

A 5.0 g portion of the above product and 1.0 g of palladium catalyst oncarbon in 50 ml of water is hydrogenated for one hour during which time2.2 pounds of hydrogen are absorbed. The mixture is filtered throughdiatomaceous earth to remove the catalyst and the filtrate isconcentrated to give the product of the example.

EXAMPLE 65,5'-[Ureylenebis(2-sulfo-1,4-phenylenecarbonylimino)]-bis[6-acetamido-1,3-naphthalenedisulfonicacid] hexasodium salt

The product of Example 5 is diluted to approximately 35 ml with water,then 4.0 ml of pyridine is added and phosgene is passed through untilthe solution is just acidic. Additional pyridine is added to make thesolution weakly basic, then it is poured with vigorous stirring into 350ml of ethyl alcohol. The resulting mixture is filtered and the productis washed with ethyl alcohol and ether to yield an orange powder. Thepowder is dissolved in 15 ml of water and the solution is made basicwith 1.5 ml of 5N sodium hydroxide, then is quickly back neutralizedwith acetic acid. The solution is poured into 200 ml of absolute ethylalcohol and heated on a steam bath to near boiling. The mixture is thencooled to room temperature and filtered. The product is washed withethyl alcohol and ether and is dried overnight at 110° C. to give theproduct of the example as a powder.

EXAMPLE 74-(4-Amino-2-sulfobenzamido)-5-hydroxy-2,7-naphthalenedisulfonic acidtrisodium salt

A 14.36 g portion of 8-amino-1-naphthol-3,6-disulfonic acid monosodiumsalt is dissolved in 40 ml of N sodium hydroxide with slight warming.This solution is slowly added to ethyl alcohol with stirring forming anoff-white solid which is filtered, washed with ethyl alcohol and etherand is dried in vacuo to yield 11.0 g of the disodium salt. A 7.26 gportion of the above product and 4.28 g of sodium acetate trihydrate isdissolved in 60 ml of water. The solution is cooled to 0° C. and 5.44 gof 4-nitro-2-sulfobenzoic anhydride is added all at once with stirring.Stirring is continued for 10 minutes and the solution is filtered. Thefiltrate is cooled in an ice bath, is acidified with 1.76 ml ofconcentrated hydrochloric acid then added to about 400 ml of ethylalcohol. The total volume is brought to 800 ml of addition of more ethylalcohol. The mixture is stirred for 15 minutes and is filtered. Theprecipitate is washed with ethyl alcohol followed by ethyl ether and isdried in vacuo at 78° C. to yield 7.3 g of4-hydroxy-5-(4-nitro-2-sulfobenzamido)-2,7-naphthalenedisulfonic acidtrisodium salt.

A 7.5 g portion of the preceding compound and 800 mg of 10% palladiumcatalyst on carbon in 125 ml of water is hydrogenated in a Parr shakeruntil no more hydrogen is absorbed. The mixture is filtered throughdiatomaceous earth and the filtrate is evaporated at 60° C. The residueobtained is dissolved in about 25 ml of ethyl alcohol and is added to400 ml of ethyl alcohol. The mixture is stirred for 1/2 hour andfiltered. The precipitate is washed with ethyl alcohol and ether, thenis dried in vacuo overnight to yield 5.6 g of the product of theexample.

EXAMPLE 84,4'-{Ureylenebis[(2sulfo-4,1-phenylenecarbonyl)imino]}-bis-[5-hydroxy-2,7-naphthalenedisulfonicacid]hexasodium salt

Phosgene gas is passed through a solution of 4.7 g of the product ofExample 7 and 1.71 g of anhydrous sodium carbonate in 30 ml of water atroom temperature until the solution becomes acidic to Congo Redindicator paper. The reaction mixture is filtered and the pH is adjustedto 7.2 with sodium carbonate, and then evaporated. The residue isdissolved in a minimum amount of water and ethanol is added toprecipitate 2.3 g of product as a solid.

EXAMPLE 9 8-(4-Amino-2-sulfobenzamido)-1,3,5-naphthalenetrisulfonic acidtetrasodium salt

To a warm solution of 23.8 g of 80.5%8-amino-1,3,5-naphthalenetrisulfonic acid in 25 ml of water and 25 ml of5N sodium hydroxide are slowly added 125 ml of absolute ethyl alcoholwith vigorous stirring. The mixture is cooled to room temperature, isfiltered and washed with 50 ml of 80% aqueous ethyl alcohol, then ethylalcohol and ether. The material is dried overnight at 110° C. to give21.0 g of 8-amino-1,3,5-naphthalenetrisulfonic acid trisodium salt as apowder.

A 4.49 g portion of the material above and 2.14 g of sodium acetatetrihydrate is dissolved in 30 ml of water. The solution is cooled to 0°C. in an ice bath and 2.72 g of 4-nitro-2-sulfobenzoic acid anhydride isadded at one time. After a few minutes the ice bath is removed and thesolution is stirred for a total of 20 minutes. The solution is filteredand the filtrate acidified with 0.88 ml of concentrated hydrochloricacid. The solution is evaporated and the residue is dissolved in 10 mlof water, then is added to 200 ml of ethyl alcohol and is stirred for1/2 hour. The precipitate is collected by filtration and washed withethyl alcohol and ether and is dried in vacuo. The dried material isdissolved in 10 ml of hot water, and 50 ml of absolute ethyl alcohol isadded with stirring for 1/2 hour. An additional 20 ml of ethyl alcoholis added with stirring continued for 10 minutes. The precipitate is thencollected by filtration, washed with ethyl alcohol and ether and driedin an Abderhalden apparatus at 110° C. overnight to yield 4.5 g ofproduct. The product is recycled through the procedure described aboveusing 1.07 g of sodium acetate trihydrate, 1.36 g of4-nitro-2-sulfobenzoic acid anhydride and 0.44 ml of concentratedhydrochloric acid, respectively. After the material is dried in vacuo aspreviously described, the dried material is dissolved in 10 ml of hotwater and 70 ml of ethyl alcohol is added slowly with stirring at roomtemperature for one hour. The precipitate is collected by filtration andis washed with 80% aqueous ethyl alcohol, ethyl alcohol and ether, thenis dried as previously described to afford 4.5 g of8-(p-nitro-2-sulfobenzamido)-1,3,5-naphthalenetrisulfonic acidtetrasodium salt as a solid.

A 3.9 g portion of the above compound and 400 mg of 10% palladiumcatalyst on carbon in 50 ml of water is hydrogenated, filtered andconcentrated as described in Example 7. The residue obtained isdissolved in 10 ml of hot water and 100 ml of absolute ethyl alcohol isadded. An oily precipitate is formed which is redissolved by addition ofmore ethyl alcohol. The solvent is then evaporated in vacuo to give 3.7g of 8-(4-amino-2-sulfobenzamido)-1,3,5-naphthalenetrisulfonic acidtetrasodium salt.

EXAMPLE 108,8'-{Ureylenebis[(2-sulfo-4,1-phenylenecarbonyl)imino]}di-1,3,5-naphthalenetrisulfonicacid octasodium salt

Phosgene is bubbled into a solution of 3.7 g of the product of Example 9and 1.21 g of anhydrous sodium carbonate in 30 ml of water at roomtemperature until the mixture is acidic to Congo red indicator. Thesolution is neutralized with sodium carbonate and then is concentrated.The residue is dissolved in 10 ml of hot water and 100 ml of absoluteethyl alcohol is added with separation of an oil. The material isredissolved and reconcentrated. The residue is again dissolved in 10 mlof hot water and 80 ml of absolute ethyl alcohol is added withseparation of an oil which does not dissolve on warming. The supernatantis decanted and ethyl alcohol is added to the oil which solidifies onbeing stirred. The solid is filtered, washed with ethyl alcohol andether and dried. The dried material is dissolved in 10 ml of hot water,and 35 ml of ethyl alcohol is added with the separation of an oil. Wateris added to effect solution, and the solution is cooled in an ice-boxwhence the oil is again separated. The mixture is heated to solution and40 ml of absolute ethyl alcohol is added again with the separation of anoil. The supernatant while hot is decanted and the residue is trituratedwith ethyl alcohol to provide a solid which is stirred for 1/2 hour. Thesolid is collected by filtration, washed with ethyl alcohol and etherand dried in vacuo to yield 3.15 g of the product of the example as ayellow solid.

EXAMPLE 11 3-(4-Amino-2-sulfobenzamido)-2-naphthoic acid

A 3.74 g portion of 2-amino-3-naphthoic acid and 4.28 g of sodiumacetate trihydrate in 30 ml of water is neutralized with 4 ml of 5Nsodium hydroxide. The solution is filtered and 30 ml of water is addedto the filtrate. The solution is cooled to 0° C. and 5.44 g of4-nitro-2-sulfobenzoic acid anhydride is added at one time to thestirred solution, then about 250 ml of water is added to insure solutionand stirring is continued for 20 minutes. The solution is filtered, thefiltrate is acidified with 1.76 ml of concentrated hydrochloric acid andheated to solubilize the formed precipitate. The solution is then cooledand allowed to stand. The solid is collected by filtration and is airdried to give 6.3 g of 3-(4-nitro-2-sulfobenzamido)-2-naphthoic acidsodium salt as a yellow solid.

To a 5.64 g portion of the above compound in 60 ml of water is added12.85 ml of N sodium hydroxide. Additional water is added to achievealmost complete solution. The solution is filtered, 500 mg of 10%palladium catalyst on carbon is added and the mixture is hydrogenated ina Parr shaker until no more hydrogen is absorbed. The mixture isfiltered through diatomaceous earth and the filter is washed with water.The filtrate and washes are combined and evaporated. The residue isdissolved in 40-50 ml of water and is acidified to pH 3. The precipitateformed is collected by filtration and is washed with water and acetone.The material is air-dried to yield 4.5 g of the product of the example.

EXAMPLE 123,3'-{Ureylenebis[(2-sulfo-4,1-phenylene)carbonylimino]}di-2-naphthoicacid tetrasodium salt

A 3.817 g portion of the product of Example 11 in 30 ml of water isneutralized to pH 7.5-8 with N sodium hydroxide, then 1.99 g ofanhydrous sodium carbonate is added and phosgene gas is passed in untilthe solution is acidic to Congo red indicator. The precipitate so formedis collected by filtration, and is washed with water, acetone and ethylether, then is dried in vacuo to yield 3.0 g of3,3'-{ureylenebis[(2-sulfo-4,1-phenylene)carbonylimino]}di-2-naphthoicacid disodium salt.

The material above is slurried in 15 ml of water and is treated with 5Nsodium hydroxide until solution is just achieved. The precipitate whichthen reappears is collected, and washed sparingly with water, thenwashed with ethyl alcohol and ether. The material is dried byconventional means to afford 1.0 g of a white solid as the product ofthe example.

EXAMPLE 13 4-(4-Amino-2-sulfobenzamido)-2,7-naphthalenedisulfonic acidtrisodium salt

A 151.7 g portion of 1-amino-3,6-naphthalenedisulfonic acid is addedwith stirring to 200 ml of water containing 44 g of sodium hydroxide.This solution is treated with activated charcoal and filtered. To thefiltrate is added ethyl alcohol with the formation of a precipitatewhich is allowed to stand. The solid is collected by filtration and setaside. More ethyl alcohol is added to the filtrate with the formation ofadditional product. This product is collected after standing and iscombined with the material previously set aside. The combined product isdissolved in water, treated with activated charcoal and isrecrystallized from ethyl alcohol. The product is collected byfiltration, washed with absolute ethyl alcohol and is dried to give1-amino-3,6-naphthalenedisulfonic acid disodium salt.

A 20.82 g portion of the preceding product and 8.2 g of sodium acetateis dissolved in 75 ml of water. The solution is cooled to about 2° C. inan ice bath and 14.4 g of 4-nitro-2-sulfobenzoic acid anhydride is addedall at once. Approximately 15 minutes after the addition is complete,the ice-bath is removed and stirring is continued for an additional 45minutes. The reaction mixture is then filtered and 150 ml of absoluteethanol is added resulting in formation of a precipitate which iscollected by filtration and washed with absolute ethanol and ether. Thematerial is dried by conventional means to give 20.70 g of4-(4-nitro-2-sulfobenzamido)-2,7-naphthalenedisulfonic acid trisodiumsalt as a powder.

The above product and 1.0 g of 10% palladium catalyst on carbon in 100ml of water are hydrogenated for about 11/2 hours. The reaction mixtureis filtered through diatomaceous earth and the filtrate is diluted toabout 500 ml with absolute ethyl alcohol. The dark solid formed iscollected by filtration and is taken up in water. Absolute ethyl alcoholis added incrementally with filtration until all of the dark materialseparates, first as a solid then as an oil. The addition of moreabsolute ethyl alcohol to the oil results in the formation of a lighttan solid which is collected by filtration and dried by conventionalmeans to yield 19.67 g of the product of the example.

EXAMPLE 144,4'-{Ureylenebis[(2-sulfo-4,1-phenylenecarbonyl)imino]}di-2,7-naphthalenedisulfonicacid hexasodium salt

A 5.68 g portion of the product of Example 13 and 4.24 g of sodiumcarbonate is dissolved in 550 ml of water. Phosgene gas is passed intothe solution until the solution is acidic to Congo red indicator withformulation of a solid. The reaction mixture is made basic with 5Nsodium hydroxide and the solid is collected by filtration. The productis recrystallized from water after treatment with activated charcoal andis oven-dried overnight to yield 3.90 g of the product of the example asa solid.

EXAMPLE 15 5-(4-Amino-2-sulfobenzamido)-4-hydroxy-2-naphthalenesulfonicacid disodium salt

An 11.1 g portion of 1-amino-8-naphthol-6-sulfonic acid is dissolved in100 ml of water and is neutralized to pH 7, then 8.56 g of sodiumacetate trihydrate is added with stirring. With stirring beingcontinued, 10.88 g of 4-nitro-2-sulfobenzoic acid anhydride is added allat once. The resulting mixture is stirred for 30 minutes and the residue(4.5 g) is filtered and set aside. The filtrate is acidified with 3.5 mlof concentrated hydrochloric acid and is concentrated. The residue isslurried in water and is filtered. The precipitate is washed with ethylalcohol and ether and is dried to yield 4.0 g of product a solid. Theresidue (4.5 g) set aside previously is recycled as above using one-halfthe amounts of sodium acetate and the mixed anhydride. To the resultingmixture is added a quantity of base sufficient to cause almost completesolution and stirring is continued for 20 minutes. The mixture isfiltered and the filtrate is acidified and concentrated to about 1/2volume with separation of a solid. This material is collected byfiltration, washed with ethyl alcohol and ether, and is dried to give2.9 g of an orange solid, giving a total yield of 6.9 g of4-hydroxy-5-(4-nitro-2-sulfobenzamido)-2-naphthalenedisulfonic aciddisodium salt.

A 5.12 g portion of the product above is stirred in 250 ml of water,then is filtered. To the filtrate is added 1.0 g of palladium catalyston charcoal, then the mixture is hydrogenated until no additionalhydrogen is absorbed. The mixture is filtered through diatomaceous earthand concentrated. Two crystallizations with ethyl alcohol from aqueoussolution yields a total of 4.4 g of the product of the example which iswashed with ethyl alcohol and ether and dried.

EXAMPLE 165,5'-{Ureylenebis[(2-sulfo-4,1-phenylenecarbonyl)imino]}bis[4-hydroxy-2-naphthalenesulfonicacid] tetrasodium salt

Phosgene gas is passed into a solution of 4.1 g of the product ofExample 15, and 1.81 g of sodium carbonate in 35 ml of water until it isacidic to Congo red indicator. The cooled solution is adjusted to pH 8by the addition, with stirring, of more sodium carbonate, then isadjusted to pH 6.5 with acetic acid. The resulting mixture is filteredand concentrated. The residue is dissolved in 40 ml of hot water then iscooled with formation of a precipitate. The product is collected byfiltration, is washed with some water, then with ethyl alcohol and etherand is dried by conventional means to yield 1.6 g of the product of theexample.

EXAMPLE 174-(4-Amino-2-sulfobenzamido)-5-hydroxy-1,7-naphthalenedisulfonic acidtrisodium salt

Under a nitrogen atmosphere, a 10.9 g portion of (87.4%)4-amino--5-hydroxy-1,7-naphthalenedisulfonic acid is dissolved in 100 mlof water and is neutralized to pH 7.0 with sodium hydroxide, then 6.45 gof sodium acetate trihydrate is added. An 8.05 g amount of4-nitro-2-sulfobenzoic acid anhydride is added at one time withstirring, and then stirring is continued for one hour. The reactionmixture is filtered, the filtrate is acidified with 2.2 ml ofconcentrated hydrochloric acid and is evaporated. The residue isdissolved in 40 ml of hot water and is filtered. To the filtrate isadded 160 ml of ethyl alcohol which causes formation of a yellow solid.After stirring for 1/2 hour, the precipitate is collected by filtration,washed with ethyl alcohol ether and dried in vacuo to yield 8.5 g ofmaterial. The filtrate is concentrated and the residue is dissolved in25 ml of hot water. The solution is filtered and 450 ml of absoluteethyl alcohol is added to the filtrate. The additional precipitatedproduct is collected, washed and dried to yield 7.3 g.

The combined product (15.8 g) is recycled with 2.23 g of sodium acetateand 4.03 g of 4-nitro-2 -sulfobenzoic acid anhydride. The reactionmixture filtrate is acidified with 1.1 ml of concentrated hydrochloricacid and is evaporated. The residue is recrystallized twice from waterwith ethyl alcohol to yield 12.9 g of product. An 11.9 g portion of thismaterial is stirred in 40 ml of N sodium hydroxide for 10 minutes, thenis acidified to pH 2 with concentrated hydrochloric acid. To thissolution is slowly added 160 ml of ethyl alcohol which causes formationof a yellow precipitate. The precipitate is collected by filtration,washed with 80% aqueous ethyl alcohol, ethyl alcohol and ether and isdried in vacuo to yield 10.5 g of5-hydroxy-4-(4-nitro-2-sulfobenzamido)-1,7-naphthalenedisulfonic acidtrisodium salt.

The product above and 1.0 g of 10% palladium catalyst on carbon in 160ml of water is hydrogentated as described in Example 7. The reactionmixture is filtered through diatomaceous earth and the filtrate isconcentrated. The residue is dissolved in 40 ml of hot water and 100 mlof absolute ethyl alcohol is added. The precipitate formed is collectedby filtration, is washed with ethyl alcohol and ether and is dried toyield 4.65 g of the product of the example.

EXAMPLE 184,4'-Ureylenebis[(2-sulfo-4,1-phenylenecarbonyl)imino]bis[5-hydroxy-1,7-naphthalenedisulfonicacid] hexasodium salt

Phosgene is bubbled into a solution of 4.0 g of the product of Example17 and 1.56 g of sodium carbonate in 35 ml of water until the mixture isacidic to Congo red indicator. The pH is adjusted to pH 8 with sodiumcarbonate and then to about pH 6.5 with glacial acetic acid. Thesolution is evaporated and the residue dissolved in 20 ml of water, then60 ml of absolute ethyl alcohol is added with separation of an oil. Theoil is collected and triturated with ethyl alcohol to yield a yellowsolid which is filtered and washed with ethanol and ether, and dried.The material is recycled as above and the product is dissolved in 20 mlof water. The solution is adjusted to pH 4.0 with concentratedhydrochloric acid and 70 ml of ethyl alcohol are added slowly withstirring. The precipitate formed is collected by filtration and iswashed with 80% aqueous ethyl alcohol, ethyl alcohol and ether, and isdried to yield 1.6 g of the product of the example as a powder.

EXAMPLE 19 8-(4-Amino-2-sulfobenzamido)-1,6-naphthalenedisulfonic acidtrisodium salt

To a solution of 25.0 g of 1-naphthylamine-3,8-disulfonic acid in 100 mlof water is added 20 ml of 5N sodium hydroxide with stirring. Themixture is warmed on a steambath and diluted with absolute ethyl alcoholto yield a crystalline precipitate. The solid is collected at roomtemperature and is washed with 75% aqueous ethyl alcohol, ethyl alcoholand ether. The product is dried in vacuo at 110° C. overnight to yield22.8 g of (87%) 8-amino-1,6-naphthalenedisulfonic acid disodium salt.

To a solution of 21.0 g of the above product and 12.3 g of sodiumacetate trihydrate in 180 ml of water at 2° C. (ice-bath) is added 16.5g of powdered 4-nitro-2-sulfobenzoic acid anhydride with rapid stirring.The mixture is stirred in the ice-bath for a total of 15 minutes, andthen is filtered. The filtrate is cooled in an ice bath and acidifiedwith 2.0 ml of concentrated hydrochloric acid, then is promptly dilutedwith 600 ml of cold ethyl alcohol. The solid so formed is collected byfiltration, washed with 85% aqueous ethyl alcohol, ethyl alcohol andether, and dried in vacuo at 90° C. to give 24.8 g of8-(4-nitro-2-sulfobenzamido)-1,6-naphthalenedisulfonic acid trisodiumsalt as a pale tan powder.

A 20.0 g portion of the preceding compound and 1.4 g of 10% palladiumcatalyst on carbon in 120 ml of distilled water is hydrogenated asdescribed in Example 7. The resulting mixture is filtered throughdiatomaceous earth and the filtrate is concentrated to about 80 ml. Thesolution is diluted with 400 ml of absolute ethyl alcohol and vigorousstirring with formation of a solid. The solid is collected by filtrationand is triturated with acetone. The material is washed with acetone anddried in vacuo at 110° C. to yield 7.0 g of the product of the exampleas a yellow solid.

EXAMPLE 208,8'-[Ureylenebis(2-sulfo-4,1-phenylenecarbonyl)imino]di-1,6-naphthalenedisulfonicacid hexasodium salt

Phosgene gas is bubbled into a solution of a 10.0 g portion of theproduct of Example 19 (prepared in the manner described) and 4.5 g ofanhydrous sodium carbonate in 100 ml of water until the solution becomesacidic to Congo red indicator. The solution is neutralized with sodiumcarbonate and the excess sodium carbonate is decomposed with aceticacid. The solution is warmed and 150 ml of ethyl alcohol is added whichcauses formation of a precipitate. The precipitate is collected byfiltration, washed with 85% ethyl alcohol, ethyl alcohol and ether anddried in vacuo at 110° C. to yield 8.8 g of the product of the exampleas a white solid.

EXAMPLE 21 4-(4-Amino-2-sulfobenzamido)-1,6-naphthalenedisulfonic acidtrisodium salt

A 50.0 g amount of (61.5%) 1-naphthylamine-4,7-disulfonic acid isdissolved in 150 ml of water and 5N sodium hydroxide is added untilbasic pH (10-11). The solution is warmed and ethyl alcohol is addeduntil a precipitate forms. The solid is collected by filtration, washedwith 85% ethyl alcohol, ethyl alcohol and ether and is dried in vacuo at110° C. to give 31.0 g of 4-amino-1,6-naphthalenedisulfonic aciddisodium salt.

A 30.0 g portion of the preceding product and 17.5 g of sodium acetatetrihydrate is dissolved in 200 ml of water and cooled to 2° C. To thissolution is added 24.0 g of 4-nitro-2-sulfobenzoic acid anhydride withvigorous stirring. The mixture is stirred in an ice bath for a total of15 minutes and the solid formed is collected and washed and dried aspreviously described to yield 33.5 g of4-(4-nitro-2-sulfobenzamido)-1,6-naphthalenedisulfonic acid trisodiumsalt as a tan powder.

A 25.0 g portion of the product above and 1.7 g of 10% palladiumcatalyst on carbon in 110 ml of water is hydrogenated as described inExample 7. The resulting mixture is filtered through diatomaceous earth,and the filtrate is diluted with 800 ml of absolute ethyl alcohol. Themixture is stirred and cooled with formation of a solid which iscollected by filtration. The solid is washed with ethyl alcohol andether and dried in vacuo at 110° C to yield 21.0 g of the product of theexample as a white solid.

EXAMPLE 224,4'-{Ureylenebis[(2-sulfo-4,1-phenylenecarbonyl)imino]}di-1,6-naphthalenedisulfonicacid hexasodium salt

Phosgene gas is bubbled into a solution of a 12.0 g portion of theproduct of Example 21 and 5.6 g of anhydrous sodium carbonate in 100 mlof water until the solution becomes acidic to Congo red indicator. Thesolution is neutralized as described in Example 20, then is warmed and300 ml of ethyl alcohol is added. An oil separates which solidifies onstanding. The solid is collected by filtration and is washed with 85%aqueous ethyl alcohol, ethyl alcohol and ether, then is dried in vacuoat 110° C. to give 11.5 g of the product of the example as a whitesolid.

EXAMPLE 23 4-(4-Amino-2-sulfobenzamido)-2,6-naphthalenedisulfonic acidtrisodium salt

A 45.0 g amount of (62.5%) 1-naphthylamine-3,7-disulfonic acid issuspended in 200 ml of water and 30 ml of 5N sodium hydroxide is addedto it. The solution is warmed and absolute ethyl alcohol is added untila cloudy precipitate forms. The solution is cooled and the solid iscollected by filtration and is washed with 85% ethyl alcohol, ethylalcohol and ether. The product is dried in vacuo at 110° C. to give 21.0g of 4-amino-2,6-naphthalenedisulfonic acid disodium salt.

To a solution of 10.58 g of the product above and 6.15 g of sodiumacetate trihydrate in 90 ml of water at 2° C. is added with vigorousstirring 8.25 g of powdered 4-nitro-2-sulfobenzoic acid anhydride. Themixture is stirred in an ice-bath for a total of 20 minutes and thenfiltered. The filtrate is cooled in an ice bath, acidified with 2.0 mlof concentrated hydrochloric acid and is diluted with 800 ml of coldethyl alcohol. The solid so formed is collected and washed and dried aspreviously described to yield 12.5 g of4-(4-nitro-2-sulfobenzamido)-2,6-naphthalenedisulfonic acid trisodiumsalt.

A mixture of 9.5 g of the preceding compound, 1.4 g of 10% palladiumcatalyst on carbon and 90 ml of water is hydrogenated as described inExample 7. The resulting mixture is filtered through diatomaceous earthand the filtrate is diluted with 600 ml of absolute ethyl alcohol and iscooled. The solid is filtered off and washed with ethyl alcohol andether, then is dried in vacuo at 110° C. to yield 7.3 g of white solidas the product of the example.

EXAMPLE 244,4'-{Ureylenebis[(2-sulfo-4,1-phenylenecarbonyl)imino]}di-2,6-naphthalenedisulfonicacid hexasodium salt

Phosgene gas is bubbled into a solution of 5.0 g of the product ofExample 23 and 2.5 g of anhydrous sodium carbonate in 50 ml of wateruntil the solution becomes acidic to Congo red indicator. The solutionis neutralized as described in Example 20, then is warmed and absoluteethyl alcohol is added until a precipitate is formed. The solid iscollected and washed with 85% aqueous ethyl alcohol, ethyl alcohol andether, then is dried in vacuo at 110° C to yield 4.3 g of the product ofthe example as a white solid.

EXAMPLE 25 4-(4-Amino-2-sulfobenzamido)-1,5-naphthalenedisulfonic acidtrisodium salt

A 14.0 g amount of 4-amino-1,5-naphthalenedisulfonic acid is suspendedin 100 ml of water and the solution is made basic with 5N sodiumhydroxide. The solution is warmed and absolute ethyl alcohol is addeduntil a solid is precipitated. The solid is collected by filtration andis washed with 85% aqueous ethyl alcohol, ethyl alcohol and ether, thendried in vacuo to yield 12.0 g of 4-amino-1,5-naphthalenedisulfonic aciddisodium salt.

To a solution of 10.5 g of the above material and 6.15 g of sodiumacetate trihydrate in 90 ml of water at 2° C. is added 8.25 g of4-nitro-2-sulfobenzoic acid anhydride with vigorous stirring. Themixture is stirred in an ice-bath for a total of 25 minutes and isfiltered. The solid is washed and dried as previously described to yield13.5 g of 4-(4-nitro-2-sulfobenzamido)-1,5-naphthalenedisulfonic acidtrisodium salt as a yellow powder.

A mixture of 9.5 g of the preceding compound, 1.4 g of 10% palladiumcatalyst on carbon and 90 ml of water is hydrogenated as described inExample 7. The resulting mixture is filtered through diatomaceous earthand the filtrate is diluted with about 800 ml of absolute ethyl alcoholand is cooled. The solid is cooled by filtration, washed with ethylalcohol and ether, then is dried in vacuo to yield 6.5 g of the productof the example as a light orange solid.

EXAMPLE 264,4'-{Ureylenebis[(2-sulfo-4,1-phenylenecarbonyl)imino]}di-1,5-naphthalenedisulfonicacid trisodium salt

Phosgene gas is bubbled into a solution of 5.0 g of the product ofExample 25 and 2.5 g of anhydrous sodium carbonate in 60 ml of wateruntil the solution becomes acidic to Congo red indicator. The solutionis neutralized as described in Example 20, then is warmed and absoluteethyl alcohol is added until a precipitate is formed. The solid iscollected and washed with 85% aqueous ethyl alcohol, ethyl alcohol andether, then is dried in vacuo at 110° C. to yield 4.0 g of the productof the example as a beige solid.

EXAMPLE 27 5-Amino-N-3,6,8-trisulfo-1-naphthylisophthalamic acidtrisodium salt

A mixture of 60.0 g of 5-nitroisophthalic acid, 300 ml of thionylchloride and 1 ml of dimethylformamide is stirred at room temperaturefor 30 minutes and then refluxed for 1 hour. The resulting clearsolution is allowed to stand 24 hours, then is evaporated to a smallvolume in vacuo. The evaporation step is then repeated with toluene andthe resulting liquid is diluted with 250 ml of hexane. The mixture isstirred and cooled until the resulting oil is solidified. The product isground to a powder and is recrystallized twice from carbon tetrachlorideto give 47.4 g of 5-nitroisophthaloyl chloride.

To a solution of 16.0 g of 8-amino-1,3,6-naphthalenetrisulfonic acidtrisodium salt and 10.0 g of sodium acetate trihydrate in 100 ml ofwater is added a solution of 8.7 g of 5-nitroisophthaloyl chloride in 75ml of diethyl ether and the mixture is stirred vigorously for 40 minutesat room temperature. The layers are separated and the aqueous phase isconcentrated in vacuo to approximately 55 ml and diluted with 100 ml ofethanol. After standing overnight, the mixture is filtered and theproduct is crystallized from 50% aqueous ethanol to give a total of 10.8g of product as the sum of several crops. The product is dried at 110°C. to give 9.3 g of 5-nitro-N-3,6,8-trisulfo-1-naphthylisophthalamicacid trisodium salt as a yellow powder.

A 9.0 g portion of the preceding product and 1.0 g of 10% palladiumcatalyst on carbon in 100 ml of water is hydrogenated for 3 hours. Themixture is filtered and the filtrate is concentrated and diluted withethanol. The mixture is filtered and the product dried at 110° C. togive 8.7 g of 5-amino-N-3,6,8-trisulfo-1-naphthylisophthalamic acidtrisodium salt as an off-white powder.

EXAMPLE 28 5,5'-Ureylenebis[N-(3,6,8-trisulfo-1-naphthyl)]isophthalamicacid octasodium salt

A solution of 3.8 g of the product of Example 27 and 1.6 g of sodiumcarbonate in 20 ml of water is treated with phosgene gas at roomtemperature until the solution is just weakly basic. The excesscarbonate is decomposed with acetic acid and the product is precipitatedwith 100 ml of ethanol. The mixture is filtered and the product is driedat 110° C. to give the product of the example as an off-white powder.

EXAMPLE 29 5-Amino-N-4,6,8-trisulfo-1-naphthylisophthalamic acid methylester trisodium salt

To a solution of 3.5 g of 8-amino-1,3,5-naphthalenetrisulfonic acidtrisodium salt and 2.24 g of sodium acetate trihydrate in 40 ml of wateris added, with stirring, 2.0 g of 3-carbomethoxy-5-nitrobenzoylchloride. Stirring is continued for one hour, then 4.0 ml of ethyl etheris added and stirring is continued for 2 hours longer. The solution isfiltered and the filtrate is concentrated. The residue is dissolved in20 ml of hot water and on addition of 20 ml of absolute ethyl alcohol aprecipitate is formed. The precipitate is mobilized with water and isfiltered and washed with 80% aqueous ethyl alcohol, ethyl alcohol andether. The filtrate is allowed to stand overnight to afford additionalproduct which is collected and washed as above. The combined product isdried by conventional means to yield 3.3 g of5-nitro-N-4,6,8-trisulfo-1-naphthylisophthalamic acid methyl estertrisodium salt.

A 3.27 g portion of the preceding product and 700 mg of 10% palladiumcatalyst on carbon in 100 ml of water is hydrogenated in a Parr shakeruntil no more hydrogen is absorbed. The resulting mixture is filteredthrough diatomaceous earth and the filtrate is evaporated. The residueis dissolved in about 15 ml of hot water and absolute ethyl alcohol isadded to a total volume of 250 ml with formation of a precipitate. Theprecipitate is collected by filtration, washed with ethyl alcohol andether and dried to yield 2.4 g of the product of the example as apowder.

EXAMPLE 30 5,5'-Ureylenebis[N-(4,6,8-trisulfo-1-naphthyl)isophthalamicacid[dimethyl ester hexasodium salt

Phosgene is bubbled into a solution of 2.0 g of the product of Example29 and 710 mg of sodium carbonate in 20 ml of water, with stirring,until acidic to Congo red indicator. The pH of the solution is adjustedto pH 6.0 with sodium carbonate, then it is filtered and concentrated.The residue is dissolved in 12 ml of hot water and 40 ml of absoluteethyl alcohol is added slowly, with stirring, for 1/2 hour. The whitesolid formed is collected and washed with 80% ethyl alcohol, ethylalcohol and ether, then is dried at 120° C. in an Abderhalden apparatusto yield 1.6 g of the product of the example.

EXAMPLE 31 5-Amino-N-4,6,8-trisulfo-1-naphthylisophthalamic acidtrisodium salt

An 8.0 g amount of 5-nitro-N-4,6,8-trisulfo-1-naphthylisophthalamic acidmethyl ester trisodium salt (prepared as described in Example 29) and122 ml of 0.1N sodium hydroxide is stirred at room temperature for 3hours. The resulting mixture is acidified to pH 2.0 with dilutehydrochloric acid and is concentrated. The residue is dissolved in 25 mlof hot water and the total volume is brought to 250 ml by the additionof absolute ethyl alcohol with stirring. The precipitate formed iscollected and washed with ethyl alcohol and ether, then is oven dried togive 7.0 g of 5-nitro-N-4,6,8-trisulfo-1-naphthylisophthalamic acidtrisodium salt.

A 6.0 g portion of the above product and 900 mg of 10% palladiumcatalyst on carbon in 130 ml of water is hydrogenated as described inExample 29. The resulting mixture is filtered through diatomaceous earthand the filtrate is concentrated. The residue is dissolved in about 20ml of hot water and ethyl alcohol is added to a total volume of 250 mlwith formation of a precipitate. The solid is collected and washed withethyl alcohol and ether and is dried by conventional means to yield 5.0g of the product of the example.

EXAMPLE 32 5,5'-Ureylenebis[N-(4,6,8-trisulfo-1-naphthyl)isophthalamicacid]octasodium salt

Phosgene gas is bubbled into a solution of 4.0 g of the product ofExample 31 and 1.46 g of sodium carbonate in 30 ml of water until acidicto Congo red indicator. The pH of the resulting mixture is adjusted topH 7.2 with sodium carbonate, then the mixture is filtered. The filtrateis concentrated and the residue is dissolved in 20 ml of hot water, then70 ml of absolute ethyl alcohol is added slowly, with stirring. Theprecipitate formed is collected by filtration, washed with 80% aqueousethyl alcohol, ethyl alcohol and ether and dried in an Abderhaldenapparatus to yield 3.0 g of the product of the example as a pink solid.

EXAMPLE 333,3'-Ureylenebis{5-[(8-hydroxy-4,6-disulfo-1-naphthenyl)aminocarbonyl]-benzoicacid}tetrasodium salt

A solution of 4.0 g of 4-amino-5-hydroxy-1,7-naphthalenedisulfonic acidin 60 ml of water is neutralized to pH 7.2 with NaOH. A 3.6 g portion ofsodium acetate trihydrate is added followed by 3.22 g of3-carbomethoxy-5-nitrobenzoyl chloride with vigorous stirring. Themixture is stirred fro 3 hours, 600 mg of sodium acetate trihydrate and500 mg of 3-carbomethoxy-5-nitrobenzoyl chloride are added and themixture is stirred for 2 more hours. A yellow solid is recovered byfiltration washed with 50% aqueous ethanol, ethanol, ether and driedgiving 2.1 g.

A 570 mg portion of this solid is suspended in 10 ml of 0.1N sodiumhydroxide and stirred overnight. A 1 ml portion of 1N sodium hydroxideis added and the mixture is stirred for 20 minutes. The mixture isacidified to pH₂ and evaporated. The residue is dissolved in 5 ml of hotwater, drilled in an ice box and the solid is collected by filtration,giving 490 mg ofN-(8-hydroxy-4,6-disulfo-1-naphthyl)-5-nitroisophthalamic acid disodiumsalt.

A 4.8 g portion of this 5-nitro compound (prepared in the mannerdescribed above) is dissolved in 250 ml of water, filtered andhydrogenated in a Parr shaker with 900 mg of 10% palladium on carbon.The mixture is filtered and the filtrate is evaporated. The residue isdissolved in 25 ml of hot water and ethanol is added to precipitate thedesired 5-amino derivative which is collected by filtration, washed withethanol and ether and dried giving 3.95 g of grey solid.

A 3.9 g portion of the 5-amino derivative is dissolved in 50 ml of waterand the pH is adjusted to 7 with sodium carbonate. A 1.63 g portion ofsodium carbonate is added and phosgene is passed through the solutionuntil it is acidic to Congo Red. The mixture is readjusted to pH7 withsodium carbonate, filtered and concentrated. The residue is dissolved in40 ml of hot water and diluted to 130 ml with ethanol. The mixture isfiltered and the solid is washed with 50% aqueous ethanol, ethanol,ether and dried giving 3.12 g of5,5'-ureylenebis[N-(8-hydroxy-4,6-disulfo-1-naphthenyl)-isophthalamicacid] hexasodium salt as a yellow solid.

This solid is dissolved in 20 ml of hot water and 35 ml of ethanol isadded with stirring. The mixture is filtered and the solid is washedwith 50% aqueous ethanol, ethanol, ether and dried. This solid isdissolved in 8 ml of water, acidified to pH2 and 35 ml of ethanol isadded. The solid is collected by filtration, washed with 80% ethanol,ethanol, ether and dried giving the final desired product.

EXAMPLE 344,4'-{Ureylenebis[(6-methyl-3,1-phenylenecarbonyl)imino]}-bis[5-hydroxy-1,7-naphthalenedisulfonicacid], tetrasodium salt

A 16 g portion of 4-amino-5-hydroxy-1,7-naphthalenedisulfonic acid in120 ml of water is adjusted to pH7.2. A 7.6 g portion of sodium acetatetrihydrate and 8.8 g of 2-methyl-5-nitrobenzoyl chloride are added withvigorous stirring. The mixture is stirred overnight at room temperature.The solid is then collected by filtration, washed with water, 80%ethanol, ethanol, ether and dried, giving 13.0 g of5-hydroxy-4-(5-nitro-o-toluamido)-1,7-naphthalenedisulfonic acid,disodium salt.

An 11.0 g portion of the above nitro derivative is 110 ml ofdimethylformamide containing 1.25 g of 10% palladium on carbon is shakenin a Parr apparatus until no further hydrogen is taken up. The reactionmixture is filtered and the filtrate is concentrated to a small volume.Ethanol is added to a total volume of 500 ml. The mixture isconcentrated to 100 ml and ether is added to precipitate a yellow solidwhich is collected by filtration, washed with ether and dried giving 7.6g of the corresponding amino derivative.

A 7.8 g portion of the amino derivative prepared as described above, and3.4 g of sodium carbonate is dissolved in 150 ml of water. Phosgene isbubbled into the solution with tap water cooling until acidic to CongoRed. The pH is adjusted to 7, and 3.4 g of sodium carbonate is added.Phosgene is passed in until the solution is acidic to Congo Red. Thesolution is concentrated, and crops are separated as they precipitate.The first crop is washed with 80% ethanol, ethanol, ether and driedgiving 1.5 g of solid which is dissolved in 15 ml of water, neutralizedwith sodium carbonate and precipitated with ether, washed with ethanoland ether and dried. A 500 mg portion of this solid is stirred with 3 mlof 1N sodium hydroxide at room temperature for 2 hours, acidified andthen filtered giving 300 mg of the desired final product.

EXAMPLE 358,8'-[Carbonylbis[imino(-2-sulfo-3,1-phenylene)carbonylimino]]-bis-1,3,6-naphthalenetrisulfonicacid octasodium salt

To a solution of 20.0 g of 8-amino-1,3,6-naphthalenetrisulfonic acidtrisodium salt and 8.75 g of sodium acetate trihydrate in 180 ml ofwater at room temperature is added 12.0 g of 3-nitrosulfobenzoicanhydride with stirring. A 400 ml portion of water is added. The mixtureis stirred at room temperature for 21/2 hours and then filtered. Thefiltrate is cooled, acidified with 2-3 ml of concentrated hydrochloricacid an diluted with ethanol. The pink solid is collected by filtration,washed with 85% ethanol, ether and dried. The filtrate is concentratedto a low volume and diluted with ethanol giving a second crop which iscollected and combined with the first crop giving a total of 27.6 g of8-(3-nitro-2-sulfobenzamido)-1,3,6-naphthalene trisulfonic acidtetrasodium salt.

A mixture of 24.0 g of the above 5-nitro derivative and 2.0 g of 10%palladium on carbon in 60 ml of water is hydrogenated on a Parr shakerat room temperature for one hour. The mixture is filtered. The filtrateis diluted with ethanol. The oil which forms is triturated with ethanolproducing a solid which is collected, washed with 85% ethanol, ethanol,ether and dried at 110° C. in vacuo giving the corresponding 3-aminoderivative.

A 1.9 g portion of this 3-amino derivative is dissolved in 40 ml ofwater. A 0.9 g portion of anhydrous sodium carbonate is added andphosgene is bubbled through the solution until it is acidic. Thesolution is neutralized with anhydrous sodium carbonate and excesssodium carbonate is destroyed by the addition of glacial acetic acid.The mixture is diluted with ethanol. The oil which separates istriturated twice with ethanol producing a solid which is collected,washed with 85% ethanol, ethanol, ether and dried. This solid isrecrystallized from water and ethanol giving 700 mg of the desired finalproduct as a white solid.

EXAMPLE 368,8'-[Carbonylbis[imino(-2-sulfo-3,1-phenylene)carbonylimino]]-bis-1,3,6-naphthalenetrisulfonicacid octasodium salt

A 6.6 g portion of 8-amino-1,3,6-naphthalenetrisulfonic acid trisodiumsalt and 2.92 g of sodium acetate trihydrate are dissolved in 180 ml ofwater at 2° C. A 4.0 g portion of 3-nitrosulfobenzoic anhydride is addedwith stirring. A 400 ml portion of water is added and the mixture isallowed to stand 21/2 hours, filtered and the filtrate is cooled,acidified with 2-3 ml of hydrochloric acid and diluted with ethanol. Thepink solid is collected by filtration, washed with 85% ethanol, etherand dried giving 2.5 g of 8-(3-nitro-2-sulfobenzamido)-1,3,6-naphthalenetrisulfonic acid, tetrasodium salt.

The above product is hydrogenated using 10% palladium on carbon ascatalyst to produce 1.9 g of8-(3-amino-2-sulfobenzamido)-1,3,6-naphthalene trisulfonic acid,tetrasodium salt as a white solid.

A 1.9 g portion of this product is dissolved in 40 ml of water and 0.9 gof anhydrous sodium carbonate is added. Phosgene is bubbled into thesolution until it becomes acidic. The solution is neutralized withsodium carbonate, and the excess sodium carbonate is destroyed withglacial acetic acid. Ethanol is added. The oil is separated andtriturated twice with ethanol. The solid is collected and washed with85% ethanol, ethanol, ether and dried. This solid is recrystallized froma mixture of water and ethanol giving 700 mg of the desired finalproduct as a white solid.

EXAMPLE 373,3'-(Carbonyldiimino)bis[5-[(8-hydroxy-3,6-disulfo-1-naphthalenyl)aminocarbonyl]]benzoicacid tetrasodium salt

A reaction mixture comprising 8.0 g of 1-naphthol-8-amine-3,6-disulfonicacid, 6.1 g of 3-carbomethoxy-5-nitrobenzoyl chloride, 6.8 g of sodiumacetate trihydrate and 80 ml of water is stirred for 3 hours, then 1.2 gof sodium acetate trihydrate and 700 mg of 3-carbomethoxy-5-nitrobenzoylchloride are added, and stirring is continued for 2 additional hours.The mixture is filtered, the filtrate is concentrated and the residue isdissolved in 60 ml of water when ethanol is added. The precipitate iscollected, washed with 80% ethanol, ethanol, ether and dried giving 11.9g of N-(8-hydroxy-3,6-disulfo-1-naphthyl)-5-nitro-isophthalamic acidmethyl ester disodium salt.

An 11.2 g portion of the above product is hydrogenated using 2.0 g of10% palladium on carbon catalyst in 150 ml of water. Filtration andprecipitation with ethanol gives 6.8 g of5-amino-N-(8-hydroxy-3,6-disulfo-1-naphthyl)-isophthalamic acid methylester disodium salt.

A mixture of 6.1 g of this product, 2.5 g of sodium carbonate and 60 mlof water is phosgenated until acidic to Congo Red. The pH is adjusted to7 with sodium carbonate, and 400 ml of ethanol is added with stirring.The solid is separated, washed with ethanol and ether and dried giving2.8 g of5,5'-ureylenebis[N(8-hydroxy-3,6-disulfo-1-naphthenyl)]isophthalamicacid dimethyl ester tetrasodium salt.

A 2.5 g portion of this product, 9 ml of 1N NaOH and 5 ml of water arestirred in a stoppered flask for one hour and then acidified to pH2. Anequal volume of ethanol is added and the solid is collected, washed with80% ethanol, ethanol, ether and dried giving 2.1 g of the desired finalproduct.

EXAMPLE 383,3'-Ureylenebis[5-(8-hydroxy-4,6-disulfo-1-naphthenyl)aminocarbonyl]benzoicacid tetrasodium salt

A mixture of 4.0 g of 1-naphthol-8-amine-3,5-disulfonic acid in 60 ml ofwater under nitrogen is neutralized to pH 7.2 with sodium hydroxide. A3.6 g portion of sodium acetate trihydrate is added followed by 3.22 gof 3-carbomethoxy-5-nitrobenzoyl chloride with vigorous stirring. Themixture is stirred for 3 hours, 600 mg of sodium acetate trihydrate and500 mg of 3-carbomethoxy-5-nitrobenzoyl chloride are added and stirringunder nitrogen is continued for 2 more hours. The solid is collected,washed with 50% aqueous ethanol, ethanol, ether and dried giving 2.1 gof solid. A 570 mg portion of this solid is suspended in 10 ml of 0.1Nsodium hydroxide and stirred overnight. A 1 ml portion of 1N sodiumhydroxide is added and the mixture is stirred for 20 minutes. Themixture is acidified to pH 2, concentrated and the residue is dissolvedin 5 ml of hot water. The solution is chilled producing 490 mg ofN-(8-hydroxy-4,6-disulfo-1-naphthenyl)-5-nitro-isophthalamic aciddisodium salt as a yellow solid.

A 4.8 g portion ofN-(8-hydroxy-4,6-disulfo-1-naphthenyl)-5-nitro-isophthalamic aciddisodium salt, prepared as described above is dissolved in 250 ml ofwater, filtered and the filtrate is hydrogenated with 900 mg of 10%palladium on carbon as catalyst. The reaction mixture is filtered. Thefiltrate is evaporated, and the residue is dissolved in 25 ml of hotwater. Ethanol is added, the solid is collected, washed with ethanol,ether and dried, giving 3.95 g of5-amino-N-(8-hydroxy-4,6-disulfo-1-naphthenyl)-isophthalamic aciddisodium salt.

A 3.9 g portion of the above amine in 50 ml of water is adjusted to pH 7with sodium carbonate. A 1.63 g portion of sodium carbonte is added andthe mixture is phosgenated until acidic to Congo Red. The pH is adjustedto 7 with sodium carbonate. The mixture is filtered. The filtrate isconcentrated and the residue is dissolved in 40 ml of hot water. Ethanolis added to a total volume of 130 ml. The solid is collected, washedwith 80% aqueous ethanol, ethanol, ether and dried giving 3.2 g of3,3'-ureylenebis[5-(8-hydroxy-4,6-disulfo-1-naphthenyl)aminocarbonyl]benzoicacid tetrasodium salt.

This material is dissolved in 20 ml of hot water. A 35 ml portion ofethanol is added with stirring. The precipitate is removed byfiltration. The filtrate yields a second crop which is collected, washedand dried giving 800 mg. This 800 mg is dissolved in 8 ml of water,acidified to pH 2 and 35 ml of ethanol is added. The resulting solid iscollected, washed with 80% ethanol, ethanol, ether and dried giving 700mg of the desired final product.

EXAMPLE 398,8'-[Carbonylbis[imino(-2-sulfo-3,1-phenylene)carbonylimino]]bis-1,4,6-naphthalenetrisulfonicacid octasodium salt

In the manner described in Example 36, treatment of 13.47 g of8-amino-1,4,6-trisulfonic acid trisodium salt, 6.1 g of sodium acetatetrihydrate and 8.23 g of 3-nitrosulfobenzoic anhydride in water gives 14g of 8-(3-nitro-2-sulfobenzamido)-1,4,6-naphthalene trisulfonic acid,tetrasodium salt.

The above is hydrogenated with 10% palladium on carbon to produce 11.3 gof 8-(3-amino-2-sulfobenzamido)-1,4,6-naphthalene trisulfonic acid is apink solid.

Treatment of this product with phosgene as previously described inExample 36 gives the title compound.

EXAMPLE 40 Preparation of Compressed Tablet

    ______________________________________                                        Ingredient               mg/Tablet                                            ______________________________________                                        Active Compound          0.5-500                                              Dibasic Calcium Phosphate N.F.                                                                         qs                                                   Starch USP               40                                                   Modified Starch          10                                                   Magnesium Stearate USP   1-5                                                  ______________________________________                                    

EXAMPLE 41 Preparation of Compressed Tablet -- Sustained Action

    ______________________________________                                        Ingredient             mg/Tablet                                              ______________________________________                                        Active Compound        0.5-500 (as acid                                       as Aluminum Lake*, Micronized                                                                        equivalent)                                            Dibasic Calcium Phosphate N.F.                                                                       qs                                                     Alginic Acid           20                                                     Starch USP             35                                                     Magnesium Stearate USP 1-10                                                   ______________________________________                                         *Complement inhibitor plus aluminum sulfate yields aluminum complement        inhibitor. Complement inhibitor content in aluminum lake ranges from          5-30%.                                                                   

EXAMPLE 42 Preparation of Hard Shell Capsule

    ______________________________________                                        Ingredient            mg/Capsule                                              ______________________________________                                        Active Compound       0.5-500                                                 Lactose, Spray Dried  qs                                                      Magnesium Stearate    1-10                                                    ______________________________________                                    

EXAMPLE 43 Preparation of Oral Liquid (Syrup)

    ______________________________________                                        Ingredient             % W/V                                                  ______________________________________                                        Active Compound        0.05-5                                                 Liquid Sugar           75.0                                                   Methyl Paraben USP     0.18                                                   Propyl Paraben USP     0.02                                                   Flavoring Agent        qs                                                     Purified Water qs ad   100.0                                                  ______________________________________                                    

EXAMPLE 44 Preparation of Oral Liquid (Elixir)

    ______________________________________                                        Ingredient             % W/V                                                  ______________________________________                                        Active Compound        0.05-5                                                 Alcohol USP            12.5                                                   Glycerin USP           45.0                                                   Syrup USP              20.0                                                   Flavoring Agent        qs                                                     Purified Water qs ad   100.0                                                  ______________________________________                                    

EXAMPLE 45 Preparation of Oral Suspension (Syrup)

    ______________________________________                                        Ingredient            % W/V                                                   ______________________________________                                        Active Compound       0.05-5                                                  as Aluminum Lake, Micronized                                                                        (acid equivalent)                                       Polysorbate 80 USP    0.1                                                     Magnesium Aluminum Silicate,                                                  Colloidal             0.3                                                     Flavoring Agent       qs                                                      Methyl Paraben USP    0.18                                                    Propyl Paraben USP    0.02                                                    Liquid Sugar          75.0                                                    Purified Water qs ad  100.0                                                   ______________________________________                                    

EXAMPLE 46 Preparation of Injectable Solution

    ______________________________________                                        Ingredient             % W/V                                                  ______________________________________                                        Active Compound        0.05-5                                                 Benzyl Alcohol N.F.    0.9                                                    Water for Injection    100.0                                                  ______________________________________                                    

EXAMPLE 47 Preparation of Injectable Oil

    ______________________________________                                        Ingredient             % W/V                                                  ______________________________________                                        Active Compound       0.05-5                                                  Benzyl Alcohol        1.5                                                     Sesame Oil qs ad      100.0                                                   ______________________________________                                    

EXAMPLE 48 Preparation of Intra-Articular Product

    ______________________________________                                        Ingredient               Amount                                               ______________________________________                                        Active Compound          2-20 mg                                              NaCl (physiological saline)                                                                            0.9%                                                 Benzyl Alcohol           0.9%                                                 Sodium Carboxymethylcellulose                                                                          1-5%                                                 pH adjusted to 5.0-7.5                                                        Water for Injection qs ad                                                                              100%                                                 ______________________________________                                    

EXAMPLE 49 Preparation of Injectable Depo Suspension

    ______________________________________                                        Ingredient            % W/V                                                   ______________________________________                                        Active Compound       0.05-5                                                                        (acid equivalent)                                       Polysorbate 80 USP    0.2                                                     Polyethylene Glycol 4000 USP                                                                        3.0                                                     Sodium Chloride USP   0.8                                                     Benzyl Alcohol N.F.   0.9                                                     HCl to pH 6-8         qs                                                      Water for Injection qs ad                                                                           100.0                                                   ______________________________________                                    

We claim:
 1. A compound of the formula: ##STR5## wherein R is SO₃ X, wherein X is alkali metal; R₁ is hydrogen; R₄, R₅ and R₆ are selected from the group consisting of hydrogen and SO₃ X, wherein X is as previously defined; R₂ is hydrogen; with the proviso that there is no R₁ or R₂ substituent when the bridgehead carbonylimino is attached at the carbon 2-position of the respective ring; with the further proviso that each naphthalene moiety must contain two or three SO₃ X substituents, wherein X is as previously defined; R₃ is SO₃ X, wherein X is as previously defined; R₇ is selected from the group consisting of hydrogen, hydroxy and SO₃ X, wherein X is as previously defined; and other pharmaceutically acceptable salts.
 2. A compound according to claim 1, wherein R, R₁, R₂, R₃, R₄, R₅ and R₆ are as previously defined; and R₇ is SO₃ X, wherein X is as previously defined.
 3. A compound according to claim 1, wherein R, R₁, R₂, R₃, R₄, R₅ and R₆ are as previously defined; and R₇ is selected from the group consisting of hydroxy and hydrogen.
 4. A compound according to claim 1, wherein R, R₁, R₃, R₄, R₅, R₆ and R₇ are as previously defined; and R₂ is hydrogen.
 5. A compound according to claim 4, wherein R, R₁, R₂, R₃, R₄, R₅ and R₆ are as previously defined; and R₇ is selected from the group consisting of hydrogen and hydroxy.
 6. A compound according to claim 4, wherein R, R₁, R₂, R₃, R₄, R₅ and R₆ are as previously defined; and R₇ is SO₃ X, wherein X is as previously defined.
 7. The compound according to claim 1, 4,4'-{ureylene-bis[(2-sulfo-4,1-phenylenecarbonyl)imino]}di-2,7-naphthalenedisulfonic acid hexasodium salt.
 8. The compound according to claim 1, 5,5'-[ureylenebis[(2-sulfo-4,1-phenylenecarbonyl)imino]]bis[4-hydroxy-2,7-naphthalenedisulfonic acid] hexasodium salt.
 9. The compound according to claim 1, 4,4'--ureylenebis[(2-sulfo-4,1-phenylenecarbonyl)imino]bis[5--hydroxy-2,7-naphthalenedisulfonic acid] hexasodium salt.
 10. The compound according to claim 1, 8,8'[ureylenebis(2-sulfo-4,1-phenylenecarbonylimino)]bis-1,6-naphthalenedisulfonic acid hexasodium salt.
 11. The compound according to claim 1, 4,4'-{ureylenebis[(2-sulfo-4,1-phenylenecarbonyl)imino]}bis-1,6-naphthalenedisulfonic acid hexasodium salt.
 12. The compound according to claim 1, 4,4'-[ureylenebis[(2-sulfo-4,1-phenylenecarbonyl)imino]]bis-2,6-naphthalenedisulfonic acid hexasodium salt.
 13. The compound according to claim 1, 4,4'-[ureylenebis[(2-sulfo-4,1-phenylenecarbonyl)imino]]bis-1,5-naphthalenedisulfonic acid trisodium salt.
 14. The compound according to claim 1, 8,8'-[carbonylbis[(imino-2-sulfo-3,1-phenylene)carbonylimino]]bis-1,3,6-naphthalenetrisulfonic acid octasodium salt.
 15. The compound according to claim 1, 8,8'-{carbonylbis[(imino-2-sulfo-3,1-phenylene)-carbonylimino]}bis-1,4,6-naphthalenetrisulfonic acid octasodium salt. 